ABSTRACT Human obesity has reached pandemic proportions, affecting all ages and socioeconomic groups, with more than 1.5 billion overweight adults and at least 500 million of them obese.1 Current medical and behavioral treatments of obesity fail to achieve their long-term therapeutic goals. Consequently, there is a pressing need for novel therapeutic approaches to obesity treatment. In this context, the possibility of a pharmacological switch (?Browning Effect?) from white to brown adipose tissue in subcutaneous fat to burn calories is highly attractive for the clinical management of obesity, as it has been convincingly demonstrated in animal models. MicroRNAs (miRNAs) represent the next wave of breakthrough therapies for human diseases. Agonists (agomirs) and antagonists (antagomirs) of miRNAs are developed to treat various human diseases. For instance, miR-122 antagomirs are in clinical testing for dyslipidemia and hepatitis C and miR-34 agomirs are in clinical testing for lung and other solid tumors. To address the critical issue of off-target effects and long-term safety of miRNA-based therapies for obesity, their targeted delivery to the cytosol of adipocytes (the action site of miRNAs) is essential. The objective of this project is to complete a working prototype for delivery and demonstrate in vivo efficacy for an innovative microRNA-based therapy of human obesity targeting subcutaneous adipocytes. We propose a therapeutic nanocarrier that is capable of molecularly targeting white adipocyte cells, delivering miRNA analogs, and triggering their transit into the cytosol using light. Our Laser Initiated Nanosyringe (LINs) miRNA delivery platform is based on a liquid perfluorocarbon (PFC) core co-loaded with a miRNA analog and a near infrared absorbing dye. The surface is stabilized using lipids to which targeting aptamers can be bound. LINs works by binding to a white adipocyte cell surface biomarker. Irradiation using short pulses of laser light causes absorption by the dye and triggers a vaporization event of the PFC core that acts like a nanosyringe, transiently opening the cell membrane and delivering miRNA directly to the cytosol. We have previously demonstrated that the LINs platform, without miRNA loading, is biocompatible and that the light needed to burst the PFC is well below ANSI safe laser fluence limits. The bursting event is gentle and highly transient, leaving the cell viable and intact. This innovative tool for nano-delivery to the cytosol on demand could have a paradigm shifting effect on localized delivery of therapeutics for obesity treatment. We will demonstrate the efficacy of LINs as a platform technology by targeting white adipose tissue (WAT) and delivering a therapeutic miRNA analog. MiRNA loading and release, confirmation of delivery to the cytosol, particle stability and cell targeting will be evaluated. In summary, the LINs platform is incredibly versatile, enabling targeting of WAT, miRNA delivery, and localized therapy. 1 The, G. B. D. O. C. et al. Global, regional and national prevalence of overweight and obesity in children and adults 1980-2013: A systematic analysis. Lancet (London, England) 384, 766-781, doi:10.1016/S0140-6736(14)60460-8 (2014). Proprietary and Confidential Property of NanoHybrids Inc